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公开(公告)号：US20130171698A1
公开(公告)日：2013-07-04
申请号：US13816826
申请日：2011-08-11
申请人： Kenji Watanabe , Hisao Moriya
发明人： Kenji Watanabe , Hisao Moriya
IPC分类号： C12P19/34
摘要： The present invention relates to a method of removing an intron contained in a gene from a eukaryotic gene, and linking only the exon sequences to prepare an expression vector comprising the linked sequences. Specifically, the invention relates to a method of preparing an expression vector containing linked exon sequences comprising amplifying exon sequences by PCR as one or more fragments from a giant fungal gene containing an intron, and linking the fragments together with a restriction enzyme-treated vector using the gap repair cloning method; a method of preparing an expression vector containing a full-length cDNA sequence by synthesizing and linking cDNA fragments from a fungal giant gene; a transformant having introduced therein an expression vector prepared by the method; a protein produced by the transformant; and a method of preparing a compound produced by the protein using the expression vector.
摘要翻译： 本发明涉及从真核基因中除去包含在基因中的内含子的方法，并且仅连接外显子序列以制备包含连接序列的表达载体。 具体地，本发明涉及一种制备含有连锁外显子序列的表达载体的方法，其包括通过PCR扩增外显子序列作为来自含有内含子的巨型真菌基因的一个或多个片段，并将所述片段与限制酶处理的载体连接，使用 间隙修复克隆方法; 通过合成和连接来自真菌巨基因的cDNA片段制备含有全长cDNA序列的表达载体的方法; 在其中引入了通过该方法制备的表达载体的转化体; 由转化体产生的蛋白质; 以及使用表达载体制备由蛋白质产生的化合物的方法。
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2.发明授权Heterologous expression of fungal polyketide synthetic gene in yeast and a method of preparing a compound produced by a protean encoded by the polyketide synthetic gene by the heterologous expression 有权酵母中真菌聚酮合成基因的异源表达和由聚酮化合物合成基因通过异源表达编码的蛋白质制备的化合物的制备方法公开(公告)号：US08980580B2
公开(公告)日：2015-03-17
申请号：US13816826
申请日：2011-08-11
申请人： Kenji Watanabe , Hisao Moriya
发明人： Kenji Watanabe , Hisao Moriya
摘要： The present invention relates to a method of removing an intron contained in a gene from a eukaryotic gene, and linking only the exon sequences to prepare an expression vector comprising the linked sequences. Specifically, the invention relates to a method of preparing an expression vector containing linked exon sequences comprising amplifying exon sequences by PCR as one or more fragments from a giant fungal gene containing an intron, and linking the fragments together with a restriction enzyme-treated vector using the gap repair cloning method; a method of preparing an expression vector containing a full-length cDNA sequence by synthesizing and linking cDNA fragments from a fungal giant gene; a transformant having introduced therein an expression vector prepared by the method; a protein produced by the transformant; and a method of preparing a compound produced by the protein using the expression vector.
摘要翻译： 本发明涉及从真核基因中除去包含在基因中的内含子的方法，并且仅连接外显子序列以制备包含连接序列的表达载体。 具体地，本发明涉及一种制备含有连锁外显子序列的表达载体的方法，其包括通过PCR扩增外显子序列作为来自含有内含子的巨型真菌基因的一个或多个片段，并将所述片段与限制酶处理的载体连接，使用 间隙修复克隆方法; 通过合成和连接来自真菌巨基因的cDNA片段制备含有全长cDNA序列的表达载体的方法; 在其中引入了通过该方法制备的表达载体的转化体; 由转化体产生的蛋白质; 以及使用表达载体制备由蛋白质产生的化合物的方法。
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